LOCUS       sequence-207382-        7349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cilia-membrane-targeted catalytic domain of inositol phosphae 
            5-phosphatase, CAAX-null.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7349)
  AUTHORS   Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, Setou M,
            Rohatgi R, Reiter JF, Ikegami K, Inoue T
  TITLE     Dynamic Remodeling of Membrane Composition Drives Cell Cycle through
            Primary Cilia Excision.
  JOURNAL   Cell. 2017 Jan 12;168(1-2):264-279.e15. doi: 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12.
  PUBMED    28086093
REFERENCE   2  (bases 1 to 7349)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "2017-01-12"; volume: "168(1-2):264-279.e15. doi"; pages: " 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12"
COMMENT     Sequence Label: 5HT6-YFP-Inpp5e
FEATURES             Location/Qualifiers
     source          1..7349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1314..1323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2676..2678,2679..2681,2682..3392)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2721..2742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2982..3001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3329..3350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4854..4975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4945..4964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4982..5437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5119..5140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5331..5350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5464..5568
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5570..5927
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5570..5761
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5593..5613)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5778..5913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5962..6756
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5962..6756
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6016..6035)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6626..6645
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6944..6963)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6988..7035
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtatca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgccacca
     1321 tggttccaga gcccggccct gtcaacagta gcaccccagc ctggggcccc gggccaccgc
     1381 cagccccagg gggcagcggc tgggtggccg ctgcgctgtg cgtggtcatc gtgctgacgg
     1441 cagccgccaa ttcgctgctg atcgcgctca tctgcacgca gcccgcgctg cgcaacacgt
     1501 ctaacttctt cctggtgtcg ctcttcacgt cggacctgat ggtgggattg gtggtgatgc
     1561 ccccagccat gctgaacgcg ctgtatgggc gctgggtact agctcgcggc ctctgtctgc
     1621 tttggaccgc cttcgacgtg atgtgctgca gcgcctccat tctcaacctc tgcctcatca
     1681 gcctggaccg ctacctgctc atcctctcgc cgctgcgcta caagctgcgc atgacagccc
     1741 cgcgtgccct ggcgctcatc ctgggtgcct ggagcctcgc cgcgctggcc tccttcctgc
     1801 ccctcttgct gggctggcat gaactgggca aagctcgaac atctgccccg ggccagtgcc
     1861 gcctattggc cagcctgcct tatgtcctcg tggcgtccgg cgtcaccttt ttcctgcctt
     1921 cgggtgccat ctgcttcacc tactgcagga tcttgctggc tgcccgcaag caggcggtgc
     1981 aagtggcctc gctcaccacg ggcacggcta cggctggcca ggccttggaa accttgcagg
     2041 tgcccaggac accacgccca gggatggagt ctgctgacag taggcgtctg accaccaagc
     2101 atagcaggaa ggccttgaag gccagcctga ccctcggcat cctgctgagc atgttctttg
     2161 tcacctggct gcccttcttt gtggccagca tagctcaggc cgtatgtgac tgcatctctc
     2221 caggtctctt cgatgtcctc acatggctgg gatactgtaa tagcaccatg aaccctatca
     2281 tctatcccct cttcatgcgg gacttcaaga gggccctggg caggttcgtg ccgtgtgtcc
     2341 actgtccccc ggagcaccgg gccagccccg cctccccctc catgtggacc tctcacagtg
     2401 gtgccaggcc aggcctcagc ctgcagcagg tgctgcccct gcctctgcca cccaactcag
     2461 attcagactc agcttcaggg ggcacctcgg gcctgcagct cacagcccag cttttgctgc
     2521 ctggagaggc gacccgggac cccccgccac ccaccagggc ccctactgtg gtcaacttct
     2581 tcgtcacaga ctctgtggag cctgagatac ggcagcatcc acttggttcc cccatgaaca
     2641 gtgctggtgg tagcgcagga ggaccggtcg ccaccatggt gagcaagggc gaggagctgt
     2701 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2761 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2821 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc
     2881 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2941 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3001 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3061 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3121 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3181 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3241 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     3301 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3361 ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt
     3421 cgaattcgtc ggatcttgca gactacaagc tccgcgcgca gccgctcctg gtgcgggccc
     3481 acagcagcct gggccccggc cggccgcgga gccccctggc ctgcgacgac tgttcccttc
     3541 gctcggccaa atcctccttc agcctcctgg cgcccatccg cagcaaggac gtccgcagca
     3601 ggagttacct ggagggcagc ctcctggcca gcggggccct gttgggggcg gatgagctgg
     3661 cccgctactt cccagaccgg aacgtggcac tcttcgtggc cacctggaac atgcagggcc
     3721 agaaggagct cccgcccagc ctggacgagt tcctgctccc agccgaggcc gactatgccc
     3781 aggacctgta tgtcatcggg gtccaggagg gctgttctga caggcgggag tgggagactc
     3841 gtctgcagga gacgctgggc ccgcactatg tgctgctgtc ctcggcggcc cacggcgtgc
     3901 tctacatgtc gctcttcatc cgcagggacc tcatctggtt ctgctcagag gtggagtgct
     3961 ccacggtgac cacacgcatc gtgtctcaga tcaagaccaa gggggccttg ggcatcagct
     4021 tcaccttttt tggcacttcc ttcctcttca tcacgtccca cttcacctca ggtgacggga
     4081 aggtggcgga gcggctgctg gactacacca ggactgtaca agccctggtc ctgcccagaa
     4141 atgtgcccga caccaacccc tatcgctcca gcgcagcgga cgtcaccacc cgcttcgatg
     4201 aggtgttctg gtttggagac ttcaacttcc gcctgagtgg cgggcgcaca gtcgtggacg
     4261 ccctcctgtg ccagggcctg gtggtggacg tgccggcgct gctgcagcac gaccagctca
     4321 tccgggagat gcggaaaggg tccatcttca agggcttcca ggagccggac atccacttcc
     4381 tcccatcata caagtttgac atcgggaagg acacgtacga cagcacctcc aagcagagga
     4441 cgccctcata cacggaccgc gtcttgtaca gaagccgcca caagggtgac atctgtcctg
     4501 tgagctactc ttcctgcccc gggatcaaga cgtccgacca ccgccctgtg tatggcctct
     4561 tccgggtgaa agtgaggccg gggcgagaca acattccgtt ggcagctggc aaatttgata
     4621 gagaactgta cttactagga attaaaagac ggatttcgaa ggagattcag aggcagcaag
     4681 cactacagag tcagaactcc agcaccatcg cctccgtttc ttgaggatcc accggatcta
     4741 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4801 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     4981 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5041 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5101 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5161 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5221 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5281 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5341 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     5401 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     5581 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5701 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5761 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5821 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5881 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     5941 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     6001 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     6061 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     6121 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     6181 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     6241 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     6301 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6361 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6421 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6481 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6541 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6601 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6661 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6721 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6781 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6841 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6901 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     6961 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     7021 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     7081 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     7141 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     7201 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7261 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7321 tccactgagc gtcagacccc gtagaaaag
//
